JP2020117440A - Nano-functional particles containing hydrophilic substance and method for producing the same - Google Patents

Abstract

To provide nano-functional particles containing a hydrophilic substance, and to provide methods for producing the same.SOLUTION: Provided are: a nano-functional particle containing a hydrophilic substance that can be obtained by immediately before spraying, mixing a liquid in which a surfactant and a good water-soluble solute are dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent, through each different channel, subsequently spraying in the state of liquid microparticles by pressurized gas, and evaporating and removing the water and the solvent; and a method for producing the same.SELECTED DRAWING: Figure 1

Description

The present invention relates to a nano-functional particle containing a hydrophilic substance and a method for producing the same.

In recent years, research on transdermal absorption technology in which an active ingredient is absorbed from the skin to induce it to directly act on the skin has been advanced. For example, a poorly water-soluble drug including a poorly water-soluble drug classified into Class 2 and Class 4 by BCS is manufactured into a S/W or S/O, and further S/O/W formulation to enhance solubility and absorbability. An improved sparingly soluble drug-surfactant complex preparation has been reported (Patent Document 1).
Further, a method for producing nanoparticles dispersed in microparticles and a nozzle for producing nanoparticles are known (Patent Document 2), and transdermal absorbability of an active ingredient, which is easily obtained by using the production method, is improved. A nano-functional particle formulation is disclosed (Patent Document 3).

By the way, vitamin C (ascorbic acid) is the most well known antioxidant, but since vitamin C has low stability and is easily decomposed in water, various vitamin C derivatives have been developed. In recent years, water-soluble vitamin C derivatives such as magnesium ascorbyl phosphate or sodium ascorbyl phosphate have been attracting attention, and cosmetics, quasi drugs, and pharmaceuticals containing these as active ingredients have been put on the market.

Further, as a functionalized preparation of these vitamin C or vitamin C derivatives, a nanoparticle dispersion liquid in which a protein and ascorbic acid or a derivative thereof are used, and its permeability is improved, and a preparation method thereof are disclosed. Reference 4).

International publication number WO2009/057808 JP, 2009-113169, A International publication number WO2016/167327 JP, 2017-066095, A

Conventional nano-functional particles relate to a nano-functional particle containing a hydrophobic substance and a method for producing the same, and a nano-functional particle containing a hydrophilic substance and a method for producing the same have not been known.

Therefore, it is an object of the present invention to obtain a nano-functional particle containing a hydrophilic substance and a method for producing the same.

As an example of the present invention, since a vitamin C derivative is relatively expensive, it is demanded that the vitamin C be functionalized by formulating it and that the vitamin C derivative be further functionalized.
However, the above-mentioned conventional nanoparticles, although mentioning the transdermal permeability, do not mention the characteristic function in the antioxidant action. Therefore, as an example of the present invention, it is an object of the present invention to obtain nano-functional particles that exhibit an antioxidant action even at a low dose or enhance the antioxidant action of an antioxidant.

As a result of intensive studies by the inventors, nano-functional particles containing a hydrophilic substance are obtained by using two specific liquids, that is, a liquid containing a good water-soluble solute and a liquid in which a hydrophilic substance is dispersed. The inventors have found that they can be obtained and completed the present invention. The nano-functional particles are useful as, for example, nano-functional particles having a good antioxidant effect.

That is, the present invention is as follows:
[1] A liquid in which a surfactant and a good water-soluble solute are dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through different flow paths, Among the microparticles containing the good water-soluble solute, which can be obtained by spraying in the state of liquid fine particles by pressurized gas after mixing immediately before spraying and vaporizing and removing the water and the solvent. A method for producing nano-functional particles containing the hydrophilic substance, which are dispersed and/or are present on the surface of the micro particles.
[2] The production method according to [1], wherein the hydrophilic substance contains at least one selected from cosmetic ingredients or pharmaceutical ingredients.
[3] The production method according to [1] or [2], wherein the hydrophilic substance is a hydrophilic substance having antioxidant ability.
[4] The production method according to [1], wherein the surfactant is sucrose fatty acid ester, sorbitan fatty acid ester, or a combination thereof.
[5] The production method according to [1], wherein the surfactant contained in the liquid in which the good water-soluble solute is dissolved in water is different from the surfactant contained in the liquid in which the hydrophilic substance is dispersed.
[6] A method for producing a particle dispersion, which comprises a step of dispersing the particles obtained by the production method according to any one of [1] to [5] in a solvent.
[7] A method for producing an external preparation for skin, including the method according to any one of [1] to [5].
[8] A method for enhancing the antioxidant action of a hydrophilic substance having antioxidant ability, using the particles produced by the method according to [3].
[9] A liquid in which a surfactant and a good water-soluble solute are dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through different flow paths, Nano-functional particles containing the hydrophilic substance, which can be obtained by spraying in a state of liquid fine particles by pressurized gas after mixing immediately before spraying and vaporizing and removing the water and the solvent.
[10] A dispersion liquid in which the particles according to [9] are dispersed in a solvent.
[11] A skin external preparation containing the particles according to [9].
[12] The particles according to [9], wherein the hydrophilic substance is a hydrophilic substance having antioxidant ability.
[13] A method for enhancing the antioxidant action of a hydrophilic substance having antioxidant ability, using the particles according to [12].
[14] A liquid in which a surfactant and a good water-soluble solute are dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through different flow paths, respectively, Production of nano-functional particles containing the hydrophilic substance, which can be obtained by spraying in a state of liquid fine particles by pressurized gas after mixing just before spraying and vaporizing and removing the water and the solvent. Method.
[15] A surfactant, a liquid in which a good water-soluble solute is dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through different flow paths, respectively, Use nano functional particles containing the hydrophilic substance, which can be obtained by spraying in the form of liquid fine particles by pressurized gas after mixing just before spraying and vaporizing and removing the water and the solvent. The method for suppressing the decomposition of hydrophilic substances.

According to the present invention, nano-functional particles containing a hydrophilic substance can be obtained by using two specific liquids, that is, a liquid containing a good water-soluble solute and a liquid in which a hydrophilic substance is dispersed. it can.

The nano-functional particles of the present invention improve, for example, the antioxidant action of the antioxidant contained therein. By using the particles, for example, an external preparation for skin in which the antioxidant action of an antioxidant substance is enhanced can be provided.

3 is an image observed with a transmission electron microscope of nanoparticles produced in Example 1. The long side of the black bar in the photograph represents the length: 200 nm. 1 is a photograph showing the appearance of the powder of Example 1 suspended in water and allowed to stand still for 1 month (left photograph: outer appearance, right photograph: outer appearance (sample placed on transparent plate, bottom Observed from the side)). It is a graph which shows the FRAP test result in Example 1 and the antioxidant action evaluation of the magnesium ascorbyl phosphate (APM) aqueous solution. It is a graph which shows the FRAP test result in Example 2 and the antioxidant action evaluation of L-ascorbic acid (Asc) aqueous solution. 3 is a graph showing the results of cell tests in the antioxidant action evaluation of Example 1 and the APM aqueous solution. 5 is a graph showing the cell test results in Example 2 and the evaluation of the antioxidant effect of the Asc aqueous solution. It is a graph which shows the cell test result in Example 1 and the antioxidant action evaluation of a high concentration APM aqueous solution.

[Method for producing nano-functional particles]
The nano-functional particles of the present invention include "a liquid in which a surfactant and a good water-soluble solute are dissolved in water (hereinafter, also referred to as an aqueous phase)" and "a surfactant in a solvent. A liquid in which a hydrophilic substance is dispersed (hereinafter, also referred to as an organic phase)" is mixed with each other through different flow paths immediately before spraying, and then, by a pressurized gas, liquid fine particles are formed. It is a nano-functional particle containing the hydrophilic substance, which can be obtained by spraying in the state and vaporizing and removing the water and the solvent.
The nano-functional particles containing the hydrophilic substance are preferably dispersed in the micro particles containing the good water-soluble solute and/or are present on the surface of the micro particles.
The above-mentioned "mixed just before spraying" means "a liquid in which a surfactant and a good water-soluble solute are dissolved in water", which were not mixed until just before spraying and flowed in different channels And "a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent" are mixed together immediately before spraying. For example, when a nozzle is used, "a surfactant and a liquid in which a good water-soluble solute is dissolved in water" and "a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent" are independent of each other. And enters the nozzle, the two liquids are mixed immediately before the spray port of the nozzle, and the mixed liquid is sprayed from the spray port of the nozzle. The nozzle used in the present invention can realize such mixing and spraying. Specifically, for example, the nanoparticle manufacturing nozzle disclosed in Patent Document 2 can be used.

The time from mixing the liquids to spraying is usually within a few seconds, for example, within 1 second, within 0.5 seconds, or within 0.2 seconds.

The pressure of the pressurized gas is preferably 0.01 to 0.5 MPa, more preferably 0.03 to 0.3 MPa, and particularly preferably 0.05 to 0.2 MPa.
In the method for producing nano-functional particles of the present invention, it is preferable that the flow rate on the aqueous phase side until mixing is relatively high (fast) as compared with the flow rate on the organic phase side until mixing.

In the method for producing nano-functional particles of the present invention, a mode can be adopted in which the aqueous phase swirls into the flow path of the organic phase and is mixed. In addition, the method for producing nano-functional particles of the present invention can adopt a mode in which the organic phase swirls into the flow channel of the aqueous phase and is mixed. Then, in the method for producing nano-functional particles according to the present invention, the organic phase and the aqueous phase flow into the flow path on the side of the spray port where the spray is made so as to swirl with each other, and mixing It is possible to adopt the mode.

In the method for producing nano-functional particles of the present invention, it is possible to adopt a mode in which the aqueous phase and the organic phase are opposed to each other (rather than swirling in) and mixed.

In the method for producing nano-functional particles of the present invention, it is possible to obtain nano-functional particles that improve the permeability of the water-soluble substance into the skin or improve the antioxidant ability of the water-soluble substance, However, it is preferable to adopt a mode in which the mixture flows by swirling into the flow path of the aqueous phase to perform mixing.

The microparticles obtained by the production method of the present invention may contain nano-functional particles in a state of being dispersed inside the microparticles and/or in a state of existing on the surface of the microparticles. Good. In particular, it is preferable that the nano-functional particles are dispersed and present on the surface of the micro particles. The particle diameter of the nano-functional particles is usually 10 to 500 nm.

When the particle size of the micro particles is 1 to 10 μm, the particle size of the nano-functional particles is usually in the range of 10 to 500 nm.

(Surfactant)
Examples of the surfactant include sucrose fatty acid esters such as sucrose stearate, sucrose palmitate, sucrose oleate, sucrose laurate, sucrose behenate, and sucrose erucate. , Sorbitan monostearate, sorbitan tristearate, sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyethylene glycol fatty acid esters, polyoxyethylene castor oil (polyethoxylated castor) oil), polyoxyethylene hydrogenated castor oil, polyoxyethylene polypropylene glycol copolymer, glycerin fatty acid ester, polyglycerin fatty acid ester and the like are preferably used.

As the sorbitan fatty acid ester, sorbitan oleate (for example, trade name: NIKKOL SO-10V, Nikko Chemicals Co., Ltd.), sorbitan palmitate (for example, trade name: NIKKOL SP-10V, Nikko Chemicals Co., Ltd.) and the like are particularly preferable. Is.
As the polyoxyethylene sorbitan fatty acid ester, POE(20) sorbitan oleate (eg, trade name: NIKKOLTO-10V, Nikko Chemicals Co., Ltd.), polysorbate 20, 40, 60, 80 and the like are particularly preferable. As the polyethylene glycol fatty acid ester, polyethylene glycol monolaurate is particularly preferable. As sucrose fatty acid ester, sucrose oleic acid ester (for example, trade name: O-1570, Mitsubishi Chemical Foods Co., Ltd.), sucrose palmitate ester (for example, trade name: P-1670, Mitsubishi Chemical Foods) Ltd.), sucrose stearates (eg trade name: S-1670, Mitsubishi Kagaku Foods Co., Ltd.), sucrose laurate esters (eg trade name: L-1695, Mitsubishi Kagaku Foods Co., Ltd.) Are suitable. As the polyoxyethylene castor oil, polyoxyethylene glycerol triricinolate 35 (Polyoxy35Castor Oil, trade name: Cremophor EL or EL-P, BSF Japan Co., Ltd.) is particularly suitable. As the polyoxyethylene hydrogenated castor oil, particularly, polyoxyethylene hydrogenated castor oil 50 and polyoxyethylene hydrogenated castor oil 60 are suitable. As the polyoxyethylene polyoxypropylene glycol copolymer, polyoxyethylene (160) polyoxypropylene (30) glycol (trade name: ADEKA Pluronic F-68, Asahi Denka Kogyo Co., Ltd.) and the like are particularly preferable. .. As the polyglycerin fatty acid ester, decaglycerin monolauric acid (Decaglyn1-L, Nikko Chemicals Co., Ltd.) and the like are preferable.

These surfactants may be used alone or in combination of two or more.
As the surfactant, it is preferable to use one in which the surfactant contained in the liquid in which a water-soluble solute is dissolved is different from the surfactant contained in the liquid in which a hydrophilic substance is dispersed. As a combination of different surfactants, the sucrose fatty acid esters (particularly sucrose oleic acid ester) can be used because the nano-functional particles of the present invention can be stably dispersed in water, and the antioxidant effect of the antioxidant is improved. ), sorbitan fatty acid esters (particularly sorbitan oleic acid ester) and polyoxyethylene sorbitan fatty acid esters (particularly polyoxyethylene sorbitan oleic acid ester), and particularly sorbitan fatty acid esters and polyoxyethylene sorbitan fatty acid A combination with an ester, particularly a combination of sorbitan fatty acid ester and a polyoxyethylene sorbitan oleic acid ester, particularly a combination of sorbitan oleic acid ester and a polyoxyethylene sorbitan oleic acid ester is preferable.

Particularly in the combination of sorbitan fatty acid ester and polyoxyethylene sorbitan fatty acid ester, it is preferable to use sorbitan fatty acid ester in the organic phase and polyoxyethylene sorbitan fatty acid ester in the aqueous phase.

Further, in addition to the point that the nano-functional particles of the present invention can be stably dispersed in water and improve the antioxidant effect of the antioxidant, different surface active agents for improving the permeability of the antioxidant into the skin As a combination of agents, a combination of sucrose fatty acid ester and sorbitan fatty acid ester, particularly a combination of sucrose oleic acid ester and sorbitan oleic acid ester is preferable.

(Good water-soluble solute)
The water-soluble substance used in the present invention may include a substance soluble in water. For example, mannitol, dextran, lactose, starch, xylitol, sorbitol, dextrin, sucrose, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, pullulan, gelatin, collagen, agar, sodium alginate, xanthan gum, Examples include polyethylene glycol and gum arabic. These good water-soluble solutes may be used alone or in combination of two or more.
The good water-soluble solute may be dissolved in water to form a liquid, and when the solvent evaporates, the solute solidifies to become a solid and may become a microparticle. The concentration of the good water-soluble solute (substance) in the liquid obtained by dissolving the good water-soluble solute in water is preferably 0.5 to 10% by mass, more preferably 1 to 5% by mass, and 2 to 4 It is particularly preferable that the content is% by mass.

(Hydrophilic substance)
The hydrophilic substance defined in the present invention means that when 1 g of solid powder is put in water and shaken vigorously at 20±0.5° C. for 5 minutes every 30 seconds, the water required for dissolution within 30 minutes is required. The amount is less than 100 ml.
As the hydrophilic substance used in the present invention, for example, a water-soluble antioxidant substance can be preferably used. For example, ascorbic acid, ascorbic acid derivative (ascorbyl magnesium phosphate, ascorbyl sodium phosphate, L-ascorbic acid-2 glucoside, 3-O-ethylascorbic acid, ascorbyl palmitate trisodium phosphate), glutathione, cysteine, lipoic acid , Phytic acid, polyphenols, riboflavin, uric acid, urobilinogen, melatonin, bilirubin, melanoidin, superoxide dismutase, glutathione peroxidase, peroxidase, catalase.

(Cosmetic ingredients)
As the hydrophilic substance used in the present invention, for example, a hydrophilic cosmetic ingredient can be preferably used. Examples of cosmetic ingredients include moisturizers, whitening agents, hair restorers, hair nourishing agents, hair growth agents, anti-hair graying agents, anti-aging agents, antioxidants, collagen synthesis promoters, anti-wrinkle agents, anti-acne agents, vitamin agents. , UV absorbers, fragrances, coloring agents, antiperspirants, cooling agents, warming agents, melanin production inhibitors, melanocyte activators, cleansing agents, slimming agents and the like.
Hydrophilic is as described above.

(Pharmaceutical ingredients)
For the hydrophilic substance used in the present invention, for example, a hydrophilic drug component can be preferably used. Examples of the pharmaceutical component include hair growth agents, hair nourishing agents, hair growth agents, antibiotics, anticancer agents, anti-inflammatory agents, antiallergic agents, hormone agents, antithrombotic agents, immunosuppressive agents, skin disease therapeutic agents, antifungal agents, nucleic acids. Drugs, anesthetics, antipyretics, analgesics, antipruritics, antiedema agents, antitussive agents, antiepileptics, antiparkinsonics, hypnotics, anxiolytics, stimulants, neuropsychiatric agents, muscle relaxants, Antidepressants, general cold drugs, autonomic nervous system agents, anticonvulsants, antiperspirants, antiperspirants, inotropic agents, antiarrhythmic agents, antiarrhythmic agents, vasoconstrictors, vasodilators, antiarrhythmic agents, antihypertensive agents, diabetes Examples include therapeutic agents, high-lipid plasma agents, respiratory stimulants, antitussives, vitamins, agents for parasitic skin diseases, homeostatic agents, polypeptides, hormones, parakeratosis inhibitors, vaccines, or emollients. it can.
Hydrophilic is as described above.

(Particles having antioxidant ability)
Antioxidant capacity, for example, refers to the ability to reduce or eliminate harmful reactions involving oxygen, such as suppressing lipid peroxidation reactions, especially in living organisms oxidative stress or reactive oxygen species (oxygen free radicals, By trapping hydroxyl radicals, superoxide anions, hydrogen peroxide, etc.), it contributes to the detoxifying reaction. The particles having an antioxidant ability refer to particles having a particle form containing or containing an antioxidant having such an action and exhibiting the above-mentioned antioxidant ability.

[Method for producing particle dispersion]
The particle dispersion of the present invention can be produced by simply adding the nano-functional particles of the present invention to a solvent and stirring the dispersion, while maintaining a good stable dispersion state. The solvent used for producing the dispersion of the present invention is not particularly limited as long as it can disperse the nano-functional particles of the present invention, and examples thereof include water, dimethyl sulfoxide, alcohols such as ethanol, and the like. A mixed solvent may be used. Water is preferred.

[Method for producing external preparation for skin]
(External skin preparation)
The particle dispersion of the present invention is suitable for use in external preparations for skin, particularly for use in external preparations for aqueous skin. For example, by adding the particle dispersion to a base material such as a thickening polysaccharide, external application is facilitated. Can be made into a drug.

[Method for enhancing the antioxidative effect of hydrophilic substances having antioxidative ability]
Using a hydrophilic substance having an antioxidant ability, by dispersing the nano-functional particles obtained by the method for producing nano-functional particles in water, a single aqueous solution of a hydrophilic substance having an antioxidant ability is obtained. It shows the same or higher antioxidant effect as compared with.

[Nano-functional particles]
The nano-functional particles of the present invention have a particle size of the nano size order (1 to 999 nm), and the micro particles have a particle size of the micro size order (1 to 999 μm). The nano-functional particles of the present invention are dispersed inside the microparticles and/or are present on the surface of the microparticles, for example dispersedly present and dispersed in the microparticles and/or Microparticles containing nano-functional particles existing on the surface of the microparticles are also referred to as composite powder. The nano-functional particles of the present invention have a particle size of 10 to 500 nm, preferably 20 to 450 nm, more preferably 30 to 400 nm, and are 0.1 to 15% by weight, preferably 0.2 to the microparticles. It is preferable to occupy 13 to 13% by weight, more preferably 0.3 to 10% by weight.

It is most preferable that the nano-functional particles of the present invention have a particle diameter of 30 to 400 nm occupying 0.3 to 10% by weight with respect to the micro particles.

The microparticles of the present invention preferably have a particle size of 1 to 10 μm.

The particle diameters of the nano-functional particles and the micro particles of the present invention are determined by DLS measurement and/or TEM observation described in the examples.

(Dispersion liquid)
The particle dispersion liquid of the present invention is a liquid in which the nano-functional particles of the present invention are stably dispersed in a solvent. The particle concentration in the dispersion is preferably 0.003 to 50 mg/mL. The production method is not limited to the method described in the above method for producing a particle dispersion. The solvent used for producing the dispersion of the present invention is not particularly limited as long as it can disperse the nano-functional particles of the present invention, and examples thereof include water, dimethyl sulfoxide, alcohols such as ethanol, and the like. A mixed solvent may be used. Water is preferred.

(External skin preparation)
The external preparation for skin of the present invention is an external preparation containing the nano-functional particles of the present invention for treating a disease by directly or indirectly applying to the skin. The external preparation for skin of the present invention is produced, for example, by dispersing the nano-functional particles of the present invention in a medium. Water or gel is mentioned as a medium. The properties are liquid (especially aqueous), ointment or gel. Since the external preparation for skin of the present application can disperse a hydrophilic substance in a medium such as water instead of dissolving it, it is excellent in the effect maintenance and the storage stability of the hydrophilic substance. The external preparation for skin of the present invention is preferably an external preparation for skin having antioxidant ability. The external preparation for skin of the present invention may contain a substance for improving the penetration of the active substance into the skin, as long as the effect of the present invention is not impaired.

Further, the external preparation for skin of the present invention can contain components that can be usually added to the external preparation for skin, as long as the effects of the present invention are not impaired. Such components include glycerin, polyhydric alcohols such as propylene glycol, liquid paraffin, squalane, higher fatty acids, oils such as higher alcohols, citric acid, organic acids such as lactic acid, caustic soda, alkalis such as triethanolamine, Cationic surfactant, amphoteric surfactant, nonionic surfactant, powder, pigment, dye, antiseptic and fungicide, resin, pH adjusting agent, antioxidant, ultraviolet absorber, chelating agent, thickener, Moisturizers, alcohol, water, fragrances, etc. are exemplified.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.
1. Preparation of Particles The reagents and equipment used for the preparation of particles in the examples are shown below.
・Reagent magnesium ascorbyl phosphate (hereinafter referred to as APM, manufactured by Ito Corporation)
L-ascorbic acid (hereinafter referred to as Asc, manufactured by Tokyo Kasei)
Sorbitan monooleate (Product name: SO-10V, referred to as SO-10V below, manufactured by Nikko Chemicals)
POE(20) sorbitan monooleate (Product name: TO-10V, hereinafter referred to as TO-10V, manufactured by Nikko Chemicals)
Mannitol (made by Tokyo Kasei)
・Apparatus spray dry nozzle: Twin jet nozzle RJ-10-TLM (Okawara Kakohki Co., Ltd.)
Liquid feed pump to spray dry nozzle: EHN-B11SH9R (manufactured by Iwaki)
Spray dry body: CL-8 (Okawara Kakoki Co., Ltd.)

Example 1 Preparation of APM-encapsulating particles
The toluene in which SO-10V was dissolved was stirred, and the APM aqueous solution was added dropwise to the mixture to prepare an emulsion in which the continuous phase was toluene, and this was used as the organic phase (charge ratio: SO-10V/toluene). /APM/water = 2/150/1/50 (w/w/w/w)). The aqueous phase was prepared by dissolving mannitol in water in which TO-10V was dissolved (charge ratio: TO-10V/water/mannitol=2/875.5/22.5 (w/w/w)). The organic phase and the aqueous phase were fed to a spray-dry nozzle using a feed pump so that the ratio was about 1:4 w/w to obtain the title particles (composite powder). The inlet temperature of the spray dry body was 180°C. The compressed nitrogen supplied to the nozzle was 0.15 MPa.
Example 2 Preparation of Asc-encapsulated particles
The toluene in which SO-10V was dissolved was stirred, and the Asc aqueous solution was added dropwise to the mixture to prepare an emulsion in which the continuous phase was toluene, and this was used as the organic phase (charge ratio: SO-10V/toluene). /APM/water = 2/150/1/50 (w/w/w/w)). The aqueous phase was prepared by dissolving mannitol in water in which TO-10V was dissolved (charge ratio: TO-10V/water/mannitol=2/875.5/22.5 (w/w/w)). The organic phase and the aqueous phase were fed to a spray-dry nozzle using a feed pump so that the ratio was about 1:4 w/w to obtain the title particles (composite powder). The inlet temperature of the spray dry body was 180°C. The compressed nitrogen supplied to the nozzle was 0.15 MPa.

2. Quantification of active ingredient (APM or Asc) content in powder and particle size measurement The APM or Asc concentration in the powder and the particle size in water obtained in Examples 1 and 2 were evaluated by HPLC and DLS measurement. did. The equipment and conditions used are as follows.
(1) HPLC measurement 0.10 g of the powder of Examples 1 and 2 was accurately weighed and added to 10 mL of water to prepare a sample. The obtained sample was subjected to HPLC under the following conditions and analyzed. The results obtained are shown in Table 1.

<HPLC conditions>
Column: TSK-gel ODS-120T (4.6 x 150 mm)
Temperature: 40℃
Detection: UV 244nm
Flow rate: 1.0 mL/min
Injection volume: 10 μL
Eluent: 20 mM ammonium acetate/5 mM tetrabutylammonium bromide/acetonitrile 80:20 (v/v) (premix)
(2) DLS measurement The powders obtained in Examples 1 and 2 were dissolved in water to prepare samples. The particle diameter in the sample was measured using a dynamic light scattering device (DLS: ZETASIZER Nano series Nano-ZS; manufactured by Malvern Instruments Ltd.). The results obtained are shown in Table 1. Table 1 shows the main peaks of the number distribution.

(3) TEM observation After dissolving the powder obtained in Example 1 in water, the nanoparticles isolated from the aqueous dispersion were subjected to a transmission electron using a centrifugal ultrafiltration container (Centrisart MWCO5,000; manufactured by Sartorius). It was observed with a microscope (TEM: manufactured by JEOL Ltd.). The observed image is shown in FIG.

From Table 1 and FIG. 1, it was confirmed that the particles obtained in the examples were nanoparticles of the nanometer order having a particle size (particle diameter) of, for example, about 80 nm.
3. Dispersion Stability in Water The powders of Examples 1 and 2 were added to water so that the APM or Asc concentration was 200 ppm to prepare dispersions. The resulting dispersion was allowed to stand for 1 month, and then its appearance was observed. No precipitate was observed and a good dispersion state was maintained (appearance of the aqueous dispersion of Example 1 left to stand for 1 month). The photograph is shown in Fig. 2). From this, it was confirmed that the nanoparticles had good dispersion stability in water.

4. Evaluation of antioxidative ability The antioxidative ability of the nanoparticles obtained in Examples 1 and 2 was evaluated by comparing with the antioxidative action of the APM or Asc aqueous solution. Antioxidant test, a test using Cell Biolabs, Inc. OxiSelect ^TM Ferric Reducing Ability of Plasma assay kit (hereinafter, referred to as FRAP test) and a test using OxiSelect ^TM intracellular antioxidant activity assay kit (hereinafter, Cell test). In the FRAP test, the absorbance was used as an index of the effect. In the cell test, the amount of decrease in fluorescence intensity with respect to the control was used as an index of effect.

(1) FRAP test: Example 1
APM was dissolved in the assay buffer to prepare a 200 ppm APM solution, and this was further diluted to prepare 100, 50, and 20 ppm APM solutions, respectively. In addition, the powder of Example 1 was added to the assay buffer to prepare an aqueous dispersion of APM-encapsulated particles of 200 ppm in terms of APM concentration, and this was further diluted to 100,50,20 ppm APM-encapsulated particle aqueous dispersion. Were prepared respectively. Next, the prepared 200, 100, 50, 20 ppm solution and dispersion were put in 96 well plates (Corning Co. 96 Well Flat Clear Bottom Black Polystyrene TC-Treated Microplates 3603) 100 μL × 6 wells, respectively, and the Reaction Reagent solution (Colorimetric) in 3 wells of them. Probe/Iron Chloride) was added 100 μL each, and assay buffer was added 100 μL each to the remaining 3 wells. The absorbance in the well prepared by the above procedure was measured by a plate reader (manufactured by TECAN), and the absorbance of the sample itself was subtracted to calculate the net absorbance. The test results are shown in FIG. From the test results, it was confirmed that the sample of Example 1 exhibited an antioxidant ability equivalent to that of the APM aqueous solution at each concentration, and did not inhibit the activity even when made into nanoparticles and exhibited an antioxidant action.

(2) FRAP test: Example 2
Asc was dissolved in the assay buffer to prepare a 200 ppm Asc solution, and this was diluted to prepare 100 and 40 ppm Asc solutions, respectively. Further, the powder of Example 2 was added to the assay buffer to prepare a 200 ppm Asc-encapsulated particle aqueous dispersion in terms of Asc concentration, and this was diluted to prepare 100,40 ppm Asc-encapsulated particle aqueous dispersion. Next, the prepared 100 and 40 ppm solution and dispersion were put in 96 well plates (Corning Co. 96 Well Flat Clear Bottom Black Polystyrene TC-Treated Microplates 3603) 100 μL x 6 wells, respectively, and Reaction Reagent solution (Colorimetric Probe/ IronChloride) was added by 100 μL, and assay buffer was added by 100 μL by the remaining 3 wells. The absorbance in the well prepared by the above procedure was measured by a plate reader (manufactured by TECAN), and the absorbance of the sample itself was subtracted to calculate the net absorbance. The test results are shown in FIG. From the test results, it was confirmed that the sample of Example 2 exhibited an antioxidant ability equivalent to that of the Asc aqueous solution at each concentration, and did not inhibit the activity even when made into nanoparticles and exhibited an antioxidant action.

(3) Cell test: Example 1
-Preparation of test plate For the cell test, normal human epidermal keratinocytes (hereinafter referred to as NHEK cells) cultured in HuMeda-KG2 medium (Kurashiki Spinning Co., Ltd.) were used. NHEK cells of 4 passages of 80×10 ⁴ cells were added to a centrifuge tube, centrifuged (300×g, 3 min), the supernatant was removed, and 8 mL of HuMeda-KG2 medium was added to each. The suspension obtained by suspending the cells by pipetting was inoculated into a 96-well plate (Corning 96 Well Flat Clear Bottom Black Polystyrene TC-Treated Microplates 3603) at 1.0 × 10 ⁴ cells/100 μL/well, and 5% CO Incubated in the presence of _{2 at} 37° C. for 24 hours. Next, a test sample was prepared by the following procedure.

・Preparation of evaluation medium
APM-containing medium of 1000 ppm was prepared by dissolving APM in HuMeda-KG2 medium, and 40, 30, 20, and 10 ppm of APM-containing medium were prepared by diluting the medium. Also, the powder of Example 1 was added to HuMeda-KG2 medium to prepare 100 ppm APM-encapsulated particle dispersion medium in terms of APM concentration, and 40,30,20,10 ppm APM-encapsulated particle dispersion medium was diluted by diluting this. Each was prepared.
・Free Radical Initiator; Preparation of 2.8% HEPES solution
FreeRadical Initiator (28 mg) was dissolved in HEPES (1000 μL) to prepare Free Radical Initiator (100x). Immediately before use, Free Radical Initiator (100x) (250 μL) was dissolved in HEPES (25 mL) to prepare a 2.8% HEPES solution.

・Test method The culture medium in each well of the test plate was removed, and DCFH-DA Probe (2x) (50uL) prepared by dissolving DCFH-DA Probe (1000x) (40μL) in HuMeda-KG2 medium (20mL) was added. well. Next, an evaluation medium (50 uL) was added to each well and incubated for 24 hours. In addition, it has been proved by WST evaluation that the evaluation medium does not affect cell survival at the sample concentration. After the incubation, the evaluation medium was removed and washed with HEPES three times. Free Radical Initiator (1x) (100uL) was added, and the fluorescence intensity was immediately measured with a plate reader. The antioxidant capacity was evaluated by the amount of decrease in fluorescence intensity when the fluorescence intensity of the control (no sample added) 60 minutes after the start of measurement was set to 0. The test results are shown in FIG. Compared with the control, in Example 1, a significant decrease in fluorescence intensity was confirmed, whereas no decrease in fluorescence intensity was observed in the APM aqueous solution. From this, it was confirmed that the sample of Example 1 exhibits significant antioxidant ability even in a very dilute concentration range of 5 ppm to 20 ppm of APM.

(4) Cell test: Example 2
-Preparation of test plate For the cell test, normal human epidermal keratinocytes (hereinafter referred to as NHEK cells) cultured in HuMeda-KG2 medium (Kurashiki Spinning Co., Ltd.) were used. NHEK cells of 4 passages of 80×10 ⁴ cells were added to a centrifuge tube, centrifuged (300×g, 3 min), the supernatant was removed, and 8 mL of HuMeda-KG2 medium was added to each. The suspension obtained by suspending the cells by pipetting was inoculated into a 96-well plate (Corning 96 Well Flat Clear Bottom Black Polystyrene TC-Treated Microplates 3603) at 1.0 × 10 ⁴ cells/100 μL/well, and 5% CO Incubated in the presence of _{2 at} 37° C. for 24 hours. Next, a test sample was prepared by the following procedure.

・Preparation of evaluation medium
Asc was dissolved in HuMeda-KG2 medium to prepare 1000 ppm Asc-containing medium, which was diluted to prepare 40, 30, 20, and 10 ppm Asc-containing medium, respectively. Further, the powder of Example 2 was added to HuMeda-KG2 medium to prepare 100 ppm Asc-encapsulated particle dispersion medium in terms of Asc concentration, and 40, 30, 20, 10 ppm Asc-encapsulated particle dispersion medium was diluted by diluting the Asc-encapsulated particle dispersion medium. Each was prepared.
・Free Radical Initiator; Preparation of 2.8% HEPES solution
FreeRadical Initiator (28 mg) was dissolved in HEPES (1000 μL) to prepare Free Radical Initiator (100x). Immediately before use, Free Radical Initiator (100x) (250 μL) was dissolved in HEPES (25 mL) to prepare a 2.8% HEPES solution.

・Test method The culture medium in each well of the test plate was removed, and DCFH-DA Probe (2x) (50uL) prepared by dissolving DCFH-DA Probe (1000x) (40μL) in HuMeda-KG2 medium (20mL) was added. well. Next, an evaluation medium (50 uL) was added to each well and incubated for 24 hours. In addition, it has been proved by WST evaluation that the evaluation medium does not affect cell survival at the sample concentration. After the incubation, the evaluation medium was removed and washed with HEPES three times. Free Radical Initiator (1x) (100uL) was added, and the fluorescence intensity was immediately measured with a plate reader. The antioxidant capacity was evaluated by the amount of decrease in fluorescence intensity when the fluorescence intensity of the control (no sample added) 60 minutes after the start of measurement was set to 0. The test results are shown in FIG. Compared with the control, in Example 2, a significant decrease in the fluorescence intensity was confirmed, whereas in the Asc aqueous solution, the decrease in the fluorescence intensity was slight. From this, it was confirmed that the sample of Example 2 showed significant antioxidant ability even in a very dilute concentration range of 5 to 20 ppm Asc.

(5) Cell Test: Comparative Examples/Preparation of Test Plate For the cell test, normal human epidermal keratinocytes (hereinafter referred to as NHEK cells) cultured in HuMeda-KG2 medium (Kurashiki Spinning Co., Ltd.) were used. NHEK cells of 4 passages of 80×10 ⁴ cells were added to a centrifuge tube, centrifuged (300×g, 3 min), the supernatant was removed, and 8 mL of HuMeda-KG2 medium was added to each. The suspension obtained by suspending the cells by pipetting was inoculated into a 96-well plate (Corning 96 Well Flat Clear Bottom Black Polystyrene TC-Treated Microplates 3603) at 1.0 × 10 ⁴ cells/100 μL/well, and 5% CO Incubated in the presence of _{2 at} 37° C. for 24 hours. Next, a test sample was prepared by the following procedure.

・Preparation of evaluation medium
APM was dissolved in HuMeda-KG2 medium to prepare a 2000 ppm APM-containing medium, which was further diluted to prepare a 400 ppm APM-containing medium. Further, the powder of Example 1 was added to HuMeda-KG2 medium to prepare 100 ppm APM-encapsulated particle dispersion medium in terms of APM concentration, and this was diluted to prepare 40,20 ppm APM-encapsulated particle dispersion medium, respectively.
・Free Radical Initiator; Preparation of 2.8% HEPES solution
FreeRadical Initiator (28 mg) was dissolved in HEPES (1000 μL) to prepare Free Radical Initiator (100x). Immediately before use, Free Radical Initiator (100x) (250 μL) was dissolved in HEPES (25 mL) to prepare a 2.8% HEPES solution.

・Test method Test plate The medium in each well was removed, and DCFH-DA Probe(2x) (50uL) prepared by dissolving DCFH-DAProbe (1000x) (40μL) in HuMeda-KG2 medium (20mL) was added to each well. Was added to. Next, an evaluation medium (50 uL) was added to each well and incubated for 24 hours. In addition, it has been proved by WST evaluation that the evaluation medium does not affect cell survival at the sample concentration. After the incubation, the evaluation medium was removed and washed with HEPES three times. Free Radical Initiator (1x) (100uL) was added, and the fluorescence intensity was immediately measured with a plate reader. The antioxidant capacity was evaluated by the amount of decrease in fluorescence intensity when the fluorescence intensity of the control (no sample added) 60 minutes after the start of measurement was set to 0. The test results are shown in FIG. Compared to the control, the fluorescence intensity decreased in all the samples, but at the same level of decrease, the concentration was 10 ppm in Example 1, which was a very dilute solution, whereas the APM aqueous solution showed 1000 ppm. The concentration was 100 times higher than that in Example 1. From this, it was shown that the antioxidant-encapsulated nanoparticles have high antioxidant ability.

The present invention is a nano-functional particle containing a hydrophilic substance, which is produced by using two specific types of liquids, and a production method thereof. The particles of the present invention can be used as an antioxidant having an enhanced antioxidant ability even with a small amount of antioxidant.

Claims (15)

A surfactant, a liquid in which a good water-soluble solute is dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through separate channels, respectively, immediately before spraying. After mixing, by pressurized gas, spraying in the state of liquid fine particles, which can be obtained by vaporizing and removing the water and the solvent, dispersed in the microparticles containing the good water-soluble solute, And/or a method for producing nano-functional particles containing the hydrophilic substance, which are present on the surface of the micro particles. The manufacturing method according to claim 1, wherein the hydrophilic substance contains at least one selected from cosmetic ingredients and pharmaceutical ingredients. The method according to claim 1 or 2, wherein the hydrophilic substance is a hydrophilic substance having an antioxidant ability. The production method according to claim 1, wherein the surfactant is sucrose fatty acid ester, sorbitan fatty acid ester, or a combination thereof. The manufacturing method according to claim 1, wherein the surfactant contained in the liquid in which the good water-soluble solute is dissolved is different from the surfactant contained in the liquid in which the hydrophilic substance is dispersed. A method for producing a particle dispersion liquid, comprising a step of dispersing the particles obtained by the production method according to claim 1 in a solvent. A method for producing an external preparation for skin, comprising the method according to claim 1. A method for enhancing the antioxidant action of a hydrophilic substance having antioxidant ability, using the particles produced by the method according to claim 3. A surfactant, a liquid in which a good water-soluble solute is dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through separate flow paths and immediately before spraying. Nano-functional particles containing the hydrophilic substance, which can be obtained by, after mixing, spraying in a state of liquid fine particles with a pressurized gas to vaporize and remove the water and the solvent. A dispersion liquid in which the particles according to claim 9 are dispersed in a solvent. A skin external preparation containing the particles according to claim 9. The particle according to claim 9, wherein the hydrophilic substance is a hydrophilic substance having an antioxidant ability. A method for enhancing the antioxidant activity of a hydrophilic substance having antioxidant capacity, using the particles according to claim 12. A surfactant, a liquid in which a good water-soluble solute is dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through separate flow paths and immediately before spraying. A method for producing nano-functional particles containing the hydrophilic substance, which can be obtained by, after mixing, spraying in a state of liquid fine particles with a pressurized gas to vaporize and remove the water and the solvent. A surfactant, a liquid in which a good water-soluble solute is dissolved in water, and a liquid in which a hydrophilic substance is dispersed in a liquid in which a surfactant is dissolved in a solvent are passed through separate flow paths and immediately before spraying. After mixing, by pressurized gas, it is sprayed in the state of liquid fine particles, which can be obtained by vaporizing and removing the water and the solvent, using the nano-functional particles containing the hydrophilic substance, hydrophilic. Method for suppressing decomposition of volatile substances.